Tricistronic overexpression of cytochrome P450_cam, putidaredoxin, and putidaredoxin reductase provides a useful cell-based catalytic system

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• 作者：Donghak Kim and Paul R. Ortiz de Montellano
• 关键词：Camphor ; Limonene ; P450 ; Tricistronic expression
• 刊名：Biotechnology Letters
• 出版年：2009
• 出版时间：September, 2009
• 年：2009
• 卷：31
• 期：9
• 页码：1427-1431
• 全文大小：282.1 KB

文摘

The catalytic turnover of cytochrome P450 _cam from Pseudomonas putida requires two auxiliary reduction partners, putidaredoxin (Pd) and putidaredoxin reductase (PdR). We report the functional expression in Escherichia coli of tricistronic constructs consisting of P450 _cam encoded by the first cistron and the auxiliary proteins, Pd and PdR by the second and the third. Transformed bacterial whole cells efficiently oxidized (1R)-(+)-camphor to 5-exo-hydroxycamphor and, interestingly, limonene to (−)-perillyl alcohol. These bioengineered E. coli cells possess a heterologous self-sufficient P450 catalytic system that may have advantages in terms of low cost and high yield for the production of fine chemicals.